Synthesis and properties of cholecystokinin-releasing peptide (monitor peptide), a 61-residue trypsin inhibitor.
A 61-residue cholecystokinin-releasing peptide (monitor peptide), which was obtained from rat pancreatic juice and found to stimulate pancreatic enzyme secretion, was recently reported to inhibit bovine trypsin and to possess epidermal growth factor (EGF)-like activities, at a concentration of about 10 nM. However, monitor peptide is structurally different from the EGF family of growth factors. To investigate whether monitor peptide contains the supposed EGF-like activities, it has been synthesized together with its [Ala23, Ala47] analog. The purified peptides, which were fully characterized by a range of methods including Cf-252 ionization mass spectrometry and enzymatic digestion to establish the locations of disulfide linkages, were shown to belong to the pancreatic secretory trypsin inhibitor family and not to the EGF family. Neither synthetic monitor peptide nor its analog were able to compete with 125I-EGF in A-431 cells or to stimulate growth of Swiss 3T3 and NRK 49F cells, up to 1 microM concentration. However, synthetic monitor peptide was as effective as the native product in the inhibition of trypsin. Replacement of the essential Arg23 in the [Ala23, Ala47]-analog led to loss of trypsin inhibition activity.
Lin
Y Z
YZ
Rockefeller University, New York, NY.
Isaac
D D
DD
Tam
J P
JP
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D004220:Disulfides / Q000737:chemistry; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000187:drug effects; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D007457:Iodine Radioisotopes; D008969:Molecular Sequence Data; D012689:Sequence Homology, Nucleic Acid; D014359:Trypsin Inhibitor, Kazal Pancreatic / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology
D004220:Disulfides; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D007457:Iodine Radioisotopes; C504705:Spink1 protein, rat; D014359:Trypsin Inhibitor, Kazal Pancreatic; D004815:Epidermal Growth Factor; D066246:ErbB Receptors